An Immunochemical Approach to Detect the Quorum Sensing-Regulated Virulence Factor 2-Heptyl-4-Quinoline N-Oxide (HQNO) Produced by Pseudomonas aeruginosa Clinical Isolates

Autor: Enrique J. Montagut, Juan Raya, M.-Teresa Martin-Gomez, Lluïsa Vilaplana, Barbara Rodriguez-Urretavizcaya, M.-Pilar Marco
Přispěvatelé: Ministerio de Ciencia e Innovación (España), Institut Català de la Salut, [Montagut EJ, Raya J, Vilaplana L, Rodriguez-Urretavizcaya B, Marco MP] Nanobiotechnology for Diagnostics (Nb4D), Department of Surfactants and Nanotechnology, Institute for Advanced Chemistry of Catalonia (IQAC) of the Spanish Council for Scientific Research (CSIC), Barcelona, Spain. CIBER de Bioingeniería, Biomateriales, y Nanomedicina (CIBER-BBN), Barcelona, Spain. [Martin-Gomez MT] Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, Spain, Vall d'Hebron Barcelona Hospital Campus
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Microbiology (medical)
fenómenos microbiológicos::interacciones microbianas::detección de quórum [FENÓMENOS Y PROCESOS]
Percepció de quòrum (Microbiologia)
Physiology
Pseudomones aeruginosa
HQNO-2-heptyl-4-quinoline N-oxide
MvfR
Virulència (Microbiologia)
Antibodies
PqsR
Bacterial Proteins
Genetics
Humans
Pseudomonas Infections
Diagnostic
Immunoassay
4-Quinolones
General Immunology and Microbiology
Ecology
Virulence factors
Virulence
Microbiological Phenomena::Microbial Interactions::Quorum Sensing [PHENOMENA AND PROCESSES]
Oxides
Cell Biology
Biological Factors::Toxins
Biological::Virulence Factors [CHEMICALS AND DRUGS]

infecciones bacterianas y micosis::infecciones bacterianas::infecciones por bacterias gramnegativas::infecciones por Pseudomonas [ENFERMEDADES]
factores biológicos::toxinas biológicas::factores de virulencia [COMPUESTOS QUÍMICOS Y DROGAS]
Quorum sensing
Infectious Diseases
Malalties bacterianes gramnegatives
Pseudomonas aeruginosa
Bacterial Infections and Mycoses::Bacterial Infections::Gram-Negative Bacterial Infections::Pseudomonas Infections [DISEASES]
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
Scientia
Popis: Understanding quorum sensing (QS) and its role in the development of pathogenesis may provide new avenues for diagnosing, surveillance, and treatment of infectious diseases. For this purpose, the availability of reliable and efficient analytical diagnostic tools suitable to specifically detect and quantify these essential QS small molecules and QS regulated virulence factors is crucial. Here, we reported the development and evaluation of antibodies and an enzyme-linked immunosorbent assay (ELISA) for HQNO (2-heptyl-4-quinoline N-oxide), a QS product of the PqsR system, which has been found to act as a major virulence factor that interferes with the growth of other microorganisms. Despite the nonimmunogenic character of HQNO, the antibodies produced showed high avidity and the microplate-based ELISA developed could detect HQNO in the low nM range. Hence, a limit of detection (LOD) of 0.60 ± 0.13 nM had been reached in Müeller Hinton (MH) broth, which was below previously reported levels using sophisticated equipment based on liquid chromatography coupled to mass spectrometry. The HQNO profile of release of different Pseudomonas aeruginosa clinical isolates analyzed using this ELISA showed significant differences depending on whether the clinical isolates belonged to patients with acute or chronic infections. These data point to the possibility of using HQNO as a specific biomarker to diagnose P. aeruginosa infections and for patient surveillance. Considering the role of HQNO in inhibiting the growth of coinfecting bacteria, the present ELISA will allow the investigation of these complex bacterial interactions underlying infections. IMPORTANCE Bacteria use quorum sensing (QS) as a communication mechanism that releases small signaling molecules which allow synchronizing a series of activities involved in the pathogenesis, such as the biosynthesis of virulence factors or the regulation of growth of other bacterial species. HQNO is a metabolite of the Pseudomonas aeruginosa-specific QS signaling molecule PQS (Pseudomonas quinolone signal). In this work, the development of highly specific antibodies and an immunochemical diagnostic technology (ELISA) for the detection and quantification of HQNO was reported. The ELISA allowed profiling of the release of HQNO by clinical bacterial isolates, showing its potential value for diagnosing and surveillance of P. aeruginosa infections. Moreover, the antibodies and the ELISA reported here may contribute to the knowledge of other underlying conditions related to the pathology, such as the role of the interactions with other bacteria of a particular microbiota environment.
This work has been funded by the Ministry of Science and Innovation (SAF2015-67476-R and RTI2018-096278-B-C21) and Fundación Marató de TV3 (TV32018-201825-30-31). The Nb4D group is a consolidated research group (Grup de Recerca) of the Generalitat de Catalunya and has support from the Departament d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (expedient: 2017 SGR 1441). CIBER-BBN is an initiative funded by the Spanish National Plan for Scientific and Technical Research and Innovation from 2013 to 2016, Iniciativa Ingenio 2010, Consolider Program, and CIBER Actions were financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. Enrique J. Montagut and Juan Raya wish to thank the FPI fellowship (BES-2016-076496 and PRE2019-087542, respectively) from the Spanish Ministry of Science and Innovation. The Custom Antibody Service (CAbS) is acknowledged for its assistance and support in the production of HQNO antibodies.
Databáze: OpenAIRE